This grant proposes to further characterize the potential role of cyclic nucleotide-independent protein kinases in eukaryotic transcription. Specifically, it is proposed to investigate properties of two of those enzymes (NI and NII) which have been purified to homogeneity in this laboratory so that a comparison can be made to other protein kinases within the cell, i.e., protein kinases found on ribonuclearprotein particles as well as those found associated with cytoplasmic ribosomes. The intranuclear localization of these (and other) protein kinases will be addressed as to whether these enzymes are specifically distributed within nucleoli/nucleoplasm; whether they are found bound to nucleosomes or to the transcription complex with RNA polymerases. In addition, attempts will be made to elucidate how many protein kinases are found within the nucleus and how these differ with regard to endogenous substrate specificity. Studies will also continue to characterize transcription in isolated nuclei and nucleoli in vitro to provide a model system for the assessment of a role of phosphorylation in eukaryotic transcription. The action of thyroid hormone (T3) on liver transcription will be addressed as a model system to try to demonstrate a specific defined role of phosphorylation by NI and/or NII in the enhanced rRNA transcription by RNA polymerase I. Similar studies are proposd regarding the number and potential localization of nuclear phosphatases which remove phosphate from acidic/basic nuclear proteins. Another enzyme which modifies nuclear proteins has also been partially purified and studies will therefore be continued to define the potential role of poly-(ADP-ribose) synthetase in eukaryotic transcription.